Literature summary for 4.1.99.12 extracted from

Singh, M.; Kumar, P.; Yadav, S.; Gautam, R.; Sharma, N.; Karthikeyan, S.
The crystal structure reveals the molecular mechanism of bifunctional 3,4-dihydroxy-2-butanone 4-phosphate synthase/GTP cyclohydrolase II (Rv1415) from Mycobacterium tuberculosis (2013), Acta Crystallogr. Sect. D, 69, 1633-1644.

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Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of His-tagged DHBPS and GCHII domains of MtbribA2 in Escherichia coli strain BL21 (DE3)	Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant bifunctional enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 20-25 mg/ml protein in 20 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM DTT, with 0.001 ml of reservoir solutioncontaining 0.1 M Na HEPES pH 7.5, 15%(w/v) PEG 8000, equilibration against 0.04 ml reservoir solution, 20°C, 2-4 days, X-ray diffraction structure determination and analysis at 3.0 A resolution	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

purified recombinant bifunctional enzyme, sitting drop vapour diffusion method, mixing of 0.001 ml of 20-25 mg/ml protein in 20 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM DTT, with 0.001 ml of reservoir solutioncontaining 0.1 M Na HEPES pH 7.5, 15%(w/v) PEG 8000, equilibration against 0.04 ml reservoir solution, 20°C, 2-4 days, X-ray diffraction structure determination and analysis at 3.0 A resolution

Mycobacterium tuberculosis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	essential for catalytic activity and in stabilization of the substrate	Mycobacterium tuberculosis
additional information	the GTP cyclohydrolase II subunit has also bound Zn2+ ions	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
D-ribulose 5-phosphate	Mycobacterium tuberculosis	-	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?
D-ribulose 5-phosphate	Mycobacterium tuberculosis H37Ra / ATCC 25177	-	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	A5U2B7	-	-
Mycobacterium tuberculosis H37Ra / ATCC 25177	A5U2B7	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged DHBPS and GCHII domains from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and gel filtration	Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-ribulose 5-phosphate	-	Mycobacterium tuberculosis	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?
D-ribulose 5-phosphate	-	Mycobacterium tuberculosis H37Ra / ATCC 25177	formate + L-3,4-dihydroxybutan-2-one 4-phosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	two conformationally different molecules of Mtb-ribA2 in the asymmetric unit that form a dimer via their GCHII domains. DHBPS and GCHII functional homodimers form a long helical-like oligomer, but the enzyme exists as a dimer in solution, isolated subunit Mtb-GCHII is a dimer while Mtb-DHBPS exists as amonomer in solution. The N-terminal DHBPS domain of Mtb-ribA2 shows an alpha + beta fold consisting of a central eight-stranded beta-sheet (beta1-beta8) surrounded by seven helices (alpha1-7alpha7)	Mycobacterium tuberculosis

Synonyms

Synonyms	Comment	Organism
DHBPS	-	Mycobacterium tuberculosis
DHBPS/GCHII	-	Mycobacterium tuberculosis
Mtb-ribA2	-	Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Mycobacterium tuberculosis

General Information

General Information	Comment	Organism
additional information	an essential bifunctional 3,4-dihydroxy-2-butanone 4-phosphate synthase/GTP cyclohydrolase II (DHBPS/GCHII) enzyme from Mycobacterium tuberculosis, Mtb-ribA2. The enzyme is composed of two conformationally different molecules of Mtb-ribA2 in the asymmetric unit that form a dimer via their GCHII domains. DHBPS and GCHII functional homodimers form a long helical-like oligomer, but the enzyme exists as a dimer in solution. The DHBPS subunit possesses an active-site loop1, a substrate channelling loop2, and a pH-sensitive loop3, structure comparisons, overview	Mycobacterium tuberculosis